P9041

Phosphodiesterase II from bovine spleen

lyophilized powder, ≥5.0 units/mg protein

• Synonym(s): 3′-Exonuclease, Spleen exonuclease, Spleen phosphodiesterase
• CAS Number: 9068-54-6
• Enzyme Commission number: 3.1.16.1 (BRENDA, IUBMB)
• EC Number: 232-965-0
• MDL number: MFCD00131884
• UNSPSC Code: 12352204
• NACRES: NA.54

Properties

• form: lyophilized powder
• specific activity: ≥5.0 units/mg protein
• mol wt: 65 kDa
• UniProt accession no.: P14099
• foreign activity: 5′-Nucleotidase ≤1% of base activity
• storage temp.: −20°C
• Gene Information: cow ... PDE2A(281971)

Description

General description

Research area: Cell signaling. Phosphodiesterase (PDE) II belongs to the PDE superfamily and includes subtypes from one to 11. They exist as a dimer and comprise of N-terminal regulatory domain, C-terminal prenylated domain, and a Zn²⁺containing catalytic domain. The bovine spleen PDE2 corresponds to a molecular weight of 65 kDa. It requires divalent cations for its enzymatic activity and has an optimum pH of 7.5.

Application

Phosphodiesterase (PDE) is any enzyme that is used to break phosphodiester bonds. It is a membrane-bound glycoprotein that is used to catalyze the hydrolysis of various nucleotide polyphosphates. Phosphodiesterase II has been used in the enzymatic digestions of purified proteins such as the P8-dGMP complex. Bovine spleen phosphodiesterase has been used to digest N-cadherin. Phosphodiesterase II from bovine is suitable for sequential and end-group analysis of poly and oligonucleotides. It may be used in kinetic studieson substrate degradation of macro-molecules.

Phosphodiesterase II from bovine spleen has been used in the:

• excision of pyridyloxobutyl (POB) base adducts from DNA
• digestion of DNA prior to cycloadenosine enrichment
• hydrolysis of DNA from blue mussels gill tissue into deoxyribonucleoside 3′-monophosphates

The product has been used in the characterization of polynucleotide chain length, base composition, and identity of terminal nucleotide. The enzyme has also been used in excision of pyridyloxobutyl (POB) base adducts from DNA. Furthermore, it has been used along with micrococcal endonuclease to hydrolyze purified DNA to 3 -nucleoside monophosphates.

Biochem/physiol Actions

Phosphodiesterase (PDE) breaks phosphodiester bonds. 3-Isobutyl-methylxanthine (IBMX) is a potential PDE2 inhibitor.

The enzyme acts on poly(A), poly(U), and poly(I). Native DNA and poly(C) are quite resistant to the action of this enzyme.

Hydrolyzes RNA, RNA-Core, 3′-alkyl- and 3′-aryl-nucleoside phosphates, and polydeoxyribonucleotides with 3′-phosphate end groups to 3′-mononucleotides.
Polynucleotides having 5′-phosphomonoester end groups are not attacked.

Unit Definition

One unit will produce acid soluble nucleotides equivalent to a ΔA₂₆₀ of 16 in 30 min at pH 6.5 at 37 °C, in a 2.0 mL reaction mixture. Substrate: RNA-Core. Actual A₂₆₀ is measured on the supernatant after precipitation of the unhydrolyzed RNA with uranyl acetate-perchloric acid reagent.

Analysis Note

Protein determined by biuret

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves